Membrane-camouflaged CRISPR/Cas9-customized nanomedicine sensitizing chemo-resistance for highly effective treatment of hepatocellular carcinoma

Drug resistance in malignant tumor is one of the main causes of clinical oncology treatment failure. Herein, a novel biomimetic nanogel system coated by Hep-G2 tumor cell membrane with co-loading of sorafenib (SF) and the CRISPR/Cas9 plasmid (coding the Cas9 protein and the sgRNA for targeting RAF-1), named as CM-NGs-CD@SF (CNCS), was developed to enhance efficacy for precise hepatocellular carcinoma (HCC) therapy and inhibit SF resistance. The camouflage of homologous targeting tumor cell membrane on the outer surface of nanogel is helpful for enhancing tumor-targeting activity (∼2.5-fold). The obtained nanogel system enables quick degradation to release CRISPR/Cas9 plasmid and SF upon triggered by the GSH overexpressed in tumor region. More importantly, the CRISPR/Cas9 plasmid could down-regulate RAF-1 expression at the genomic level, meanwhile to synergize with the SF-activated RAF/MEK/ERK pathway, thus significantly enhancing the sensitivity of SF while precisely initiating apoptosis. The in vitro and in vivo results confirmed that this CNCS system could greatly improve the transfection efficiency of the CRISPR/Cas9 plasmid and effectively reducing the expression of RAF-1 genes/proteins. The transcriptome further reveals the therapeutic mechanisms by which CNCS inhibit tumor growth and splenomegaly. This multi-functionalized nanogel design based on the combination of GSH smart response and CRISPR/Cas9 technology provides an important reference for the treatment of HCC.